CRACK HEALING IN CONCRETE USING VARIOUS BIO INFLUENCED

SELF-HEALING TECHNIQUES

By
M. Basit Ehsan Khan
(NUST201260956MSCEE15212F)
Master of Science
in
Structural Engineering

NUST Institute of Civil Engineering (NICE)

School of Civil and Environmental Engineering (SCEE)
National University of Sciences and Technology (NUST) Islamabad, Pakistan

This is to certify that the

thesis titled

CRACK HEALING IN CONCRETE USING VARIOUS BIO INFLUENCED SELFHEALING TECHNIQUES

submitted by

M. Basit Ehsan Khan

NUST201260956MSCEE15212F

has been accepted towards the partial fulfillment

of the requirements for the degree
of
MASTER OF SCIENCE
in
STRUCTURAL ENGINEERING

_____________________________
Dr. Wasim Khaliq
Associate Professor
NUST Institute of Civil Engineering (NICE)
School of Civil and Environmental Engineering (SCEE)
National University of Sciences and Technology (NUST)
Islamabad, Pakistan

ii

DEDICATED
TO
MY PARENTS AND SIBLINGS

iii

ACKNOWLEDGEMENTS
In the name of Allah, the most merciful, the most compassionate all praises be to Allah, the lord
of the worlds and prayers and Peace be upon Muhammad his servant and messenger.
The completion of this project was only possible due to unlimited blessings of almighty Allah
and collaboration of many people, to whom I wish to express my gratitude.
First and foremost, i would like to thank my beloved parents and siblings for their unconditional
love and support throughout my life and for strengthening me to chase my dreams.
I would like to express my profound gratitude to my supervisor Dr. Wasim Khaliq, associate
professor, NUST institute of civil engineering, for his guidance and support throughout this
project and especially for his confidence in me. I am grateful to him for motivating me along this
arduous course. He inspired and encouraged me to be the best version of me. A special thanks
for his countless hours of reflecting, reading, encouraging and most of all patience throughout
the entire process.
There are number of people without whom this thesis might not have been written, and to whom
I am gratefully indebted. I was fortunate to have an outstanding committee. I owe my sincere
gratitude to Dr. Khaliq-ur-Rasheed Kiyani, Dr. Shaukat Ali Khan, and Dr. Syed Ali Rizwan, who
were more than generous with their expertise and precious time.
I am grateful to my colleagues especially Mr. Hammad Anis Khan, Mr. Waqas Javaid and Mr.
Tahir for their sincere help and guidance during this thesis work. I am also thankful to the
laboratory staff of structures lab, for their assistance and help during experimentation.

iv

ABSTRACT
Concrete is most widely used engineering material in construction industry due to its strength,
durability and low cost as compared to other construction materials. However, crack formation
and progression under tensile stress is a major weakness of concrete. These cracks can make
concrete vulnerable to deleterious environment due to ingress of harmful compounds, hence
compromising its durability resulting in deterioration of concrete. For a durable and good
structural concrete control over microcracks development and propagation of these cracks is
necessary and crack healing can be helpful in mitigation of development and propagation of
cracks in concrete.
Self-healing concrete involves the crack repairing in concrete by use of compounds, bacteria and
resins, added in concrete during the mixing stage. Bio concrete or bio influenced self-healing
concrete is a product which has the ability to fill the cracks by producing chemical products from
intentionally incorporated micro-organism in the concrete. In this kind of concrete microorganisms along with a precursor compound are introduced in the concrete during mixing phase.
When cracks are produced, water seeps in the cracks and activates the already present dormant
micro-organisms. These micro-organisms later produce minerals to fill the cracks hence
inhibiting strength loss and increasing durability of concrete.
An experimental program was conducted to study the self-healing mechanism in concrete under
various conditions. Bacteria were introduced in concrete by different techniques such as directly
incorporating in mix, by immobilizing it in light weight aggregate and in combination with
graphite nano platelets. In all the techniques, calcium lactate was used as an organic precursor
and replaced 5% of cement. Specimens were made for each mix to compare the change in
compressive strength of each mix. In addition to that, concrete specimens were also subjected to
pre cracking at 3,7,14 and 28 days to determine the crack healing efficiency of each mix.
Results show that bacteria immobilized in graphite nano platelets were more effective in samples
pre cracked at 3 and 7 days while bacteria immobilized in light weight aggregates were more
efficient in samples pre cracked on 14 and 28 days. In addition, results of compressive strength
depict that self-healing concrete made with light weight aggregate incorporating immobilized
bacteria had significant enhancement in compressive strength of concrete.
v

TABLE OF CONTENTS
CONTENT

Page No.

ACKNOWLEDGEMENTS ........................................................................................................... iv
ABSTRACT .................................................................................................................................... v
TABLE OF CONTENTS ............................................................................................................... vi
LIST OF FIG.URES .................................................................................................................... viii
LIST OF TABLES .......................................................................................................................... x
INTRODUCTION .......................................................................................................................... 1
1. General ................................................................................................................................. 1
2. Objectives ............................................................................................................................ 4
3. Organization of the report .................................................................................................... 5
LITERATURE REVIEW ............................................................................................................... 6
1. General ................................................................................................................................. 6
2. Previous studies on bio influenced self-healing practice ..................................................... 6
3. Summary ............................................................................................................................ 19
EXPERIMENTAL PROGRAM ................................................................................................... 21
1. General ............................................................................................................................... 21
2. Materials ............................................................................................................................ 22
2.1

Micro-organism .............................................................................................................. 22

2.2

Graphite nano platelets ................................................................................................... 23

2.3 Light weight aggregate ..................................................................................................... 24

3. Mix proportion ................................................................................................................... 25
4. Test specimens ................................................................................................................... 26
5. Test procedure .................................................................................................................... 27
RESULTS AND DISCUSSIONS ................................................................................................. 31
vi

1. General ............................................................................................................................... 31
1.1 Self-healing analysis ......................................................................................................... 31
1.2 Microstructure analysis ...................................................................................................... 37
1.3 X- Ray diffraction analysis ................................................................................................ 40
1.4 Compressive strength analysis ........................................................................................... 41
CONCLUSIONS AND RECOMMENDATIONS ....................................................................... 44
1. General ............................................................................................................................... 44
2. Conclusions ........................................................................................................................ 44
3. Recommendations .............................................................................................................. 44
REFERENCES ............................................................................................................................. 46

vii

LIST OF FIG.URES
Fig. 1.1 Process of self-healing of crack through bacteria 3
Fig. 2.1 Compressive strength of cement based mortar under various self-healing conditions 7
Fig. 2.2 Difference in 7 and 28 day compressive strength of mortar in various self-healing
conditions ... 7
Fig. 2.3 Effect of bacteria (Shewanella) addition on mortar strength 8
Fig. 2.4 Development of compressive strength of control and organic compound incorporated
mortar . 9
Fig. 2.5 Water permeability results of concrete samples against various crack widths .. 10
Fig. 2.6 Water Permeability results of concrete samples against various crack widths .. 11
Fig. 2.7 Comparison of change in transmission time under various crack filling techniques at
crack depth 10 mm ... 11
Fig. 2.8 Comparison of conductivity in different carrier compounds ..... 12
Fig. 2.9 Strength regain percentage for various mix combinations ..... 12
Fig. 2.10 Threshold values of maximum crack healed 13
Fig. 2.11 Comparison of 28 day strength of mixes with and without incorporation of LWA .... 14
Fig. 2.12 Water permeability coefficient of samples in different curing conditions ... 15
Fig. 2.13 Healing ratio of samples in various curing conditions ..... 15
Fig. 2.14 Flexural strength at various replacement levels of polyurethane foam wastes 16
Fig. 2.15 Effect of GNP on strength of cement based mortar ........ 17
Fig. 2.16 Comparison of flexural strength in PU, GNP and LWA incorporated concrete ......18
Fig. 2.17 Effect of various bacteria on compressive strength of self-healing concrete ... 19
viii

Fig. 3.1 Bacteria solution in a cylinder 23

Fig. 3.2 Graphite nano platelets ....... 24
Fig. 3.3 Light weight aggregates ..... 25
Fig. 3.4 Casting of samples .. 27
Fig. 3.5 Capping of cylinders, compression machine and cylinders after compressive strength
tests .......... 28
Fig. 3.6 Crack measuring microscope and measurement of crack by it .......... 29
Fig. 3.7 Marking of cracks on pre-cracked specimens to ensure that multiple reading are taken
from the same point ......... 29
Fig. 3.8 Closer view of cracks showing self-healing process with calcium carbonate
formation....... 30
Fig. 4.1 Crack width measurement by crack measuring microscope ...... 31
Fig. 4.2 Crack healing in specimens pre-cracked at 3 days ..... 33
Fig. 4.3 Crack healing in specimens pre-cracked at 7 days ..... 34
Fig. 4.4 Crack healing in specimens pre-cracked at 14 days ... 35
Fig. 4.5 Crack healing in specimens pre-cracked at 28 days ... 36
Fig. 4.6 Scanning electron microscope analysis of 7 days pre-cracked samples .... 38
Fig. 4.7 Scanning electron microscope (SEM) analysis of 28 days pre-cracked samples ... 40
Fig. 4.8 XRD analysis of healing compound produced in the cracks .. 41
Fig. 4.9 Compressive strength development with different bacteria incorporation techniques 42

ix

LIST OF TABLES
Table 3.1 Description of test specimens . 21
Table 3.2 Properties of GNP .. 24
Table 3.3 Mix design of different sets of specimens .. 26

CHAPTER 1
INTRODUCTION
1. General
Self-healing concrete is a product which has the ability to fill the cracks by producing chemical
products from intentionally incorporated bacteria, compounds or resins present in it. One of these
methods of self-healing, through the production of minerals by using mineral producing bacteria
in the concrete, is known as bio-influenced self-healing concrete or bio concrete. Bio concrete is
a product which heals the cracks produced in it by producing the mineral compounds to fill the
cracks. During earlier studies bacterial solution was sprayed on the cracks formed in a structure
which was a less effective and impractical procedure. However, in recent years bacteria along
with organic mineral precursor compound are incorporated in the concrete during the mixing
phase.
Concrete is most widely used engineering material in construction industry due to its strength,
durability and low cost as compared to other construction materials. However, it has certain
drawbacks as well. A major drawback of concrete is its low tensile strength which makes it
susceptible to progression and coalescence in microcracks resulting in low strength and
durability. These tensile stresses can be due to tensile loading plastic shrinkage and expansive
reactions (Mehta and Monteiro, 2006). This liability to cracking not only results in strength
reduction of concrete but also makes concrete vulnerable to deleterious environment. Cracking in
concrete is a major concern as these microcracks permit the ingress of harmful chemicals in
concrete structures. This entry of harmful chemicals may result in concrete deterioration and can
also cause corrosion of steel reinforcement (Reinhardt and Jooss, 2003). This corrosion leads to
increase in crack width and length ultimately resulting in loss of strength and stiffness of
concrete structures (Hewlett, 2003). To get a more durable structural concrete propagation of
cracks must be controlled. This deterioration of both concrete and reinforcement results in high
maintenance cost. According to report of Federal Highway Administration (2001) United States
of America spends 4 billion dollars annually in terms of direct cost of maintenance of concrete
highway bridges. De Rooij and Schlangen (2011) stated that UK spends 45 % of its annual
construction cost on maintenance of already constructed concrete structures.
1

These drawbacks made scientists and researchers to look for an optimum solution to increase the
durability of concrete by finding a solution to counter these cracks propagation in concrete.
Scientists has made use of several materials, such as epoxy systems, acrylic resins and silicone
based polymers to control this problem. However, most of these materials were found to be noncompatible with concrete, hazardous to environment and expensive (Vekariya and Pitroda,
2013).
As a result of current studies, bio concrete or bio influenced self-healing concrete is emerging as
a viable solution the tensile crack formation in concrete structures. Bio concrete involves the
incorporation of two component healing system in the concrete mixture, comprising of suitable
bacteria and calcium based nutrient compound. Autonomous healing through this process
increases the structure durability and on the other hand reduces the manual maintenance required
for structures through the process of bio-mineralization. This self-healing technique reduces the
use of environmental unfriendly repair materials and increases compressive strength of concrete,
hence saving the environment and economy (Vekariya and Pitroda, 2013).
Generally, bacteria are incorporated in dormant form along with the nutrient compound which
stays inactive until cracks are produced allowing water penetration in the concrete. Bacteria
become active when come in contact with water and begin to feed on nutrient compound.
Calcium carbonate is produced as the result of metabolic action of bacteria on calcium lactate
and seals the crack, restricting the water penetration in concrete hence increasing durability of
concrete.

(a)- Bacteria incorporated in concrete mix

(b)- Crack produced in concrete due to tensile stresses

(c)- Water is deliberately entered in the cracks

(d)- CaCO3 is produced which heals the cracks

Fig. 1.1 Process of self-healing of crack through bacteria
Self-healing in concrete is dependent on certain parameters including dissolved inorganic carbon,
pH of concrete, nucleation sites and presence of calcium ions (Hammes et al., 2003). Additional
factors include In addition, other variables such as type of bacteria, their varying concentrations,
various curing procedures and material used for incorporation of bacteria also contribute towards
efficient self-healing of concrete
Many species of Bacillus family have been proved to meet these standards and are used in this
process. These bacteria can form spores when face harsh environments and become dormant.
However, when concrete cracks and water is introduced in the cracks, alkalinity of concrete
decreases and these bacteria become active again and start producing calcium carbonate by
feeding on the nutrient compound.

In addition to the choice of suitable bacteria, selection of nutrient compound is also of great
significance as bacteria yield calcium carbonate by feeding on it. Most commonly two type of
nutrient compounds are used i.e. calcium lactate (CaC6H10O6) and urea (CO(NH2)2). However,
while using urea calcium rich environment is also required for the development of calcium
carbonate.

2. Objectives
The main objective of this study is to investigate the self-healing response of concrete when
bacteria are introduced using various techniques. This will be done in the following manner:

Introduction of bacteria samples in the concrete mix without any carrier compound and
study its efficiency in self-healing process.

Comparing the self-healing process in concrete through different carrier compounds and
identifying the optimum technique of bacteria introduction.

Studying the viability of graphite nanoplatelets (GNP) as a new carrier compound for
bacteria in concrete.

Studying the effect of using bacteria and nutrient compound (in this case calcium lactate)
on the strength of concrete.

Three different techniques were used for incorporation of bacteria in concrete which involve
direct incorporation of bacteria during mixing, by incorporation of light weight aggregate
containing immobilized bacteria and by incorporation of graphite nano platelets containing
immobilized bacteria. Out of these, effect of introducing bacteria through graphite nano platelets
(GNP) has not been studied before. Compressive strength of concrete was measured by
subjecting the concrete specimen to compression tests whereas; self-healing process was
monitored by visual inspection and SEM analysis of pre-cracked samples.
To accomplish the above mentioned objectives several tasks were undertaken. Literature review
was first conducted. Testing and data acquisition setup was designed to acquire the required
results. Samples configuration was determined and specimen were made on basis of this
configuration. Compressive tests were conducted to determine the compressive strength. For precracking, specimens were subjected to compressive load carefully. These pre-cracked samples
were then placed in curing tank and crack width was measured on regular intervals to determine
4

the efficiency of self-healing process. On basis of data gathered comparison was made between
efficiency of various self-healing techniques and their effect on compressive strength of concrete
specimen.

3. Organization of the report

Chapter 1 is an introductory chapter about bio influenced self-healing concrete, objective of the
study and thesis overview. A brief literature review on bio-concrete is discussed in chapter 2.
Chapter 3 represents the procedure and materials of test setup, the testing facility and
construction of specimens. The testing apparatus is also elaborated in Chapter 3. Chapter 4
discusses the tests carried out, the observations, test results and evaluation of test results. The
conclusions based on findings of this research and recommendations for further studies are
presented in Chapter 5.

CHAPTER 2
LITERATURE REVIEW
1. General
Bio-influenced self-healing concrete is expected to be a viable solution to the environmental
concerns of carbon dioxide production by cement industries as well as durability of concrete
structures. Due to these advantages it has been the focus of several research works and studies
during the past few years. This chapter contains notable studies carried out on bio-influenced
self-healing concrete. These research works have taken into account different variables
contributing towards self-healing of concrete. Some of these variables are type of bacteria, precursor compound and techniques used for incorporation of bacteria.

2. Previous studies on bio influenced self-healing practice

A number of research studies have been carried out on bio influenced self-healing process. Few
of important studies are presented here:
Ramachandran et al. (2001) studied the role of micro-organisms in remediation of cracks and
their effect on compressive strength in mortar samples. Micro-organisms used in this research
were Bacillus Pasteurii. For this purpose Portland cement mortar beams and beams of
dimensions 25 x 25 x 150 mm and 50 x 50 x 50 mm respectively were made and 3.175 mm wide
cracks with varying depths were produced. Cracks were filled with sand mixed with bacterial
specimen of 3 x 108 cells/cm3. Samples were placed in solutions of urea and CaCl2 for 28 days.
Compressive tests were performed to find out the compressive strength of samples. In addition to
compressive tests, SEM and XRD tests were also conducted to find out the crack healing
efficiency of bacteria. Results showed that at lower concentrations B.Pasteurii increased the
compressive strength of mortar sample. Microbiological remediation is more effective in shallow
cracks as compared to deep cracks. Fig. 2.1 depicts the results of compressive tests as observed
by Ramachandran (2001) at various concentrations. Fig. 2.2 indicates that living B.Pasteurii at
the concentration of 7.6 x 103 cells/ml showed maximum strength at the age of 28 days.

70
Control

Strength (MPa)

65

Live B.Pasteurii Conc 7.6x10E3

Live B.Pasteurii Conc 7.6x10E7
Killed B.Pasteurii Conc 7.6 x10E3

60

Killed B.Pasteurii Conc 7.6 x10E7

55
50
45
40
7

28
Days

Fig. 2.1 Compressive strength of cement based mortar under various self-healing conditions
25
Living B.Pasteurii & P. Aernglonsa
Living B.Pastuerii
Killed B.Pasteurii & P. Aernglonsa
Killed B.Pasteurii

Strength diff (MPa)

20
15
10
5
0
7.60E+03

7.60E+05

7.60E+07

-5
-10

Concentration (cells/cm3)

Fig. 2.2 Difference in 7 and 28 day compressive strength of mortar in various self-healing
conditions
Ghosh et al. (2005) studied the effect of anaerobic bacteria of Shewanella species and
Escherichia Coli on the compressive strength of cement mortar. For this purpose mortar cubes
specimens of 70.6 mm were studied. Different cell concentrations of both bacteria were used in
these samples ranging from zero to 107 per ml. It was observed through results that Shewanella
bacteria had a positive effect on compressive strength at a cell concentration of 105 per ml and
increased compressive strength by 25%. However, by increasing its concentration further caused
a decrease in compressive strength. On the other hand, addition of E.Coli bacteria had no
7

significant effect on the compressive strength of mortar samples. In addition to compressive

strength, SEM and MIP tests were also conducted on the samples. Results of these tests showed
an improvement in pore size distribution at Shewanella cell concentration of 105 per ml. increase
in compressive size was attributed to this improved pore size distribution in samples.
Improvement in compressive strength at various concentrations throughout the curing period can
be seen in the Fig. 2.3.

Avg compressive strength (MPa)

30
25
20
15

0
10
1.00E+02
1.00E+03
1.00E+04
1.00E+05
1.00E+06
1.00E+07

10
5
0
3

14

28

Curing (Days)

Fig. 2.3 Effect of bacteria (Shewanella) addition on mortar strength

Jonkers et al. (2010) studied the direct incorporation of bacteria along with nutrient compound in
the concrete. Two different species of bacillus family were added (Bacillus Cohnii and Bacillus
Pseudofirmus) in the concrete together with four different nutrient compounds including calcium
acetate, yeast, peptone and calcium lactate. Compression tests were conducted and results
exhibited that calcium acetate and yeast lowered the strength of concrete to half to that of control
specimen. Peptone addition resulted in complete detrimental effect on concrete. Only the
incorporation of calcium lactate increased compressive strength of concrete in 28 days cured
samples as shown in Fig. 2.4. Crack surfaces of both control and bacterial concrete samples was
inspected and a difference was noted among them in 7 day cured samples but surfaces of both
samples were not substantially different in 28 days cured samples. Copious amount of particles
with size 20-80m was found on crack surface of 7 days cured bacterial sample which was

absent in 28 days cured samples. This along with results from most probable number (MPN)
techniques proved that viability of bacterial spores decreases significantly over time.
70

Control
Bacteria
peptone
Yeast extract
CaAcetate
CaLactate

60

Strenght (MPa)

50
40
30
20
10
0
0

10
15
Curing (Days)

20

25

30

Fig. 2.4 Development of compressive strength of control and organic compound incorporated
mortar
De Belie and De Muynck (2008) studied the effect of crack healing using bio deposition.
Bacillus Sphaericus culture was used in this study. In order to study it standardized cracks of 0.3
mm were created by incorporating thin copper plates in concrete and pulling them out one day
after casting or by performing split tensile tests on samples wrapped in fiber reinforced polymer.
Cracked samples were cured in nutrient solution containing CaCl2 or Ca(NO3)2. Bacteria were
immobilized in silica gel and introduced in the concrete samples for protection. Visual
inspection, ultrasound testing and water permeability tests were conducted. Visual inspection
and ultrasound tests confirmed crack healing up to 0.3 mm in width and 10 mm in depth. Water
permeability tests showed that 0.6 mm wide cracks were healed through the process of bio
deposition. In addition to that water permeability test also depicts that epoxy, BS+sol-gel+CaCl2
and BS+sol-gel+Ca(NO3)2 were most efficient in reducing water permeability as shown in Fig.
2.5.

1.8E-09
Untreated
BS+ CaCl2
Epoxy
Grout
BS+sol-gel+Ca(NO3)2
Sol-gel
BS+sol-gel+CaCl2

1.6E-09
1.4E-09
K (m/s)

1.2E-09
1.0E-09
8.0E-10
6.0E-10
4.0E-10
2.0E-10
0.0E+00
0

0.2

0.4
0.6
Crack width (mm)

0.8

Fig. 2.5 Water permeability results of concrete samples against various crack widths
Van Tittelboom et al. (2010) investigated the efficiency of bacteria to repair cracks in concrete in
comparison with traditional crack repairing methods. During this study water permeability tests,
ultrasound transmission measurements and visual examinations were conducted to determine the
crack healing efficiency of various crack repairing techniques. Cracks were created by means of
split tensile methods and by creating grooves using copper wire. Crack treatment with
B.Sphaericus immobilized in silica gel showed increased ultrasonic pulse velocity. Fig.s 2.6 and
2.7 show the results of water permeability and ultrasonic pulse velocity tests respectively. As it
can be seen, bacterial solution incorporated in silica gel and treated with CaCl2 had the least
value of water permeability coefficient (K) and shows great reduction in water permeability as
compared to untreated samples. In the same way, results for ultrasonic pulse velocity tests at the
crack width of 10 mm show that maximum reduction in transmission time is observed in samples
containing bacterial solution incorporated in silica gel and treated with CaCl2. Through this
increase in ultrasonic pulse velocity together with visual examination and water permeability
tests, it was concluded that enhanced crack repair can be obtained by treating crack with
biological mix containing B.Sphaericus culture incorporated in silica gel along with calcium
source.

10

K (m/s)

1.E+00
1.E-01
1.E-02
1.E-03
1.E-04
1.E-05
1.E-06
1.E-07
1.E-08
1.E-09
1.E-10
1.E-11
1.E-12

BS in sol-gel+CaCl2
Untreated
grout
Epoxy
BS+CaCl2
Sol-gel+BS+CaCl2
BS in sol-gel + Ca(NO3)2
BS in sol-gel+ Ca(CH3COO)2

0.2

0.4

0.6
Crack width

0.8

Fig. 2.6 Water Permeability results of concrete samples against various crack widths

Change in transmission time (s)

1
0.5
0
-0.5
-1
-1.5
-2

Untreated
Grout
Epoxy
Sol-gel
BS+CaCl2
Sol-gel+BS+CaCl2
BS in sol-gel+CaCl2
BS in sol-gel + Ca(NO3)2
BS in sol-gel+ Ca(CH3COO)2

Fig. 2.7 Comparison of change in transmission time under various crack filling techniques at
crack depth 10 mm
Wang et al. (2012) studied the possibility of using silica gel (SG) and polyurethane (PU) as a
carrier for bacteria into the concrete mix. Bacteria used for this purpose was B.Sphaericus along
with urea as the nutrient precursor compound. In order to identify the efficiency of bacteria, two
different suspensions were prepared and incorporated with SG and PU. The bacterial suspensions
11

containing bacteria in alive form were represented by BS and those containing dead bacteria
were named BSA. Bacteria together with nutrient and other compounds were encapsulated in
glass tubes. When cracking occurred, glass tubes broke with it causing the encapsulate solution
to release and heal the cracks. Fig. 2.8 and 2.9 demonstrates the results of conductivity tests at
different intervals during the curing period and the strength regain observed in the prepared
specimens.
25
Polyurethane + Bacteria

20
15
10
5
0
160

180

200

220
Time (hrs)

240

260

280

Fig. 2.8 Comparison of conductivity in different carrier compounds

70
60
Strength regain (%)

Conductivity (ms/cm)

Silica Gel + Bacteria

50
40
30
20
10
0
Refernce

SG+BSA

SG+BS
PU+BSA
Mix combinations

PU+BS

Fig. 2.9 Strength regain percentage for various mix combinations

12

It can be seen that specimen having bacteria incorporated in silica gel showed better conductivity
as compared to those incorporated in polyurethane. Results showed calcium carbonate
precipitation of 25% by mass in silica gel as compared to 10% by mass in polyurethane. Fig. 2.9
shows the results of strength regain in both samples. Strength regain in case of silica gel was
relatively low i.e. 5% as compared to 60% strength regain in polyurethane. Based on these
conclusions polyurethane was nominated a better option as bacterial carrier.
Wiktor and Jonkers (2011) carried out tests on cement mortar samples. During their research
study bacteria were made inactive by heating them at 80oC for 30 minutes. Bacteria along with
nutrient precursor were protected by infusing them in light weight aggregates (LWA).
Reinforced mortar specimen were made by using ordinary Portland cement, fine aggregate and
LWA either soaked with bacteria and calcium lactate in case of bacterial specimen or nonimpregnated in case of control specimen. After 56 days of curing, cracks were created and were
re-cured. Fig. 2.10 shows threshold crack width at various healing days.
0.5
Control

Threshold crack width (mm)

0.45

Bacteria

0.4
0.35
0.3
0.25
0.2
0.15
0.1
0.05
0
0

20

40

60
80
Healing time (Days)

100

120

Fig. 2.10 Threshold values of maximum crack healed

In the light of observations made by them, they concluded that width of completely healed cracks
was more than double in bacterial specimen (46 mm) as compared to that in control specimen
(18 mm).

13

Sierra-Beltran and Jonkers (2012) discussed the effect and use of bio-based mortar for concrete
repair. Initially, in this research four different types of ECC minerals were studied and depending
upon their drying shrinkage and mechanical properties two of them were selected for further
testing as bio based mortar. In these selected two material fillers, either sand or limestone was
replaced with LWA having bacteria on them. Calcium lactate was used as nutrient precursor
compound. The incorporation of healing agent had a negative impact on flexural strength and
deflection but caused increase in compressive strength. However, all the test results exhibited
that mortars having self-healing agent lied in the acceptable limit described in European standard
EN 1504-3
60
Mix 2 (FA,BFS,Sand)
Mix 2 (FA,BFS,Sand)

50
Strength (MPa)

Mix 4 (FA,LP)
40

Mix 4 (FA,LP)

30
20
10
0
Compressive strength

Flexural strength

Fig. 2.11 Comparison of 28 day strength of mixes with and without incorporation of LWA
Wang et al. (2014) conducted research study on self-healing efficiency of microencapsulated
bacteria. During this study B.Sphaericus was used as a healing agent. Six different series of
samples were prepared containing control specimen, specimens containing nutrients, with
microcapsules, having both microcapsules and nutrients, containing nutrients and 3%
microencapsulated

bacterial

spores

(NCS3%)

and

containing

nutrients

and

5%

microencapsulated bacterial spores (NCS5%). Cracks were produced in these specimens and
after that five different incubation techniques were used to check role of water in self-healing
process. These incubation techniques included placing specimen in air conditioned room,

14

immersion in water, immersion in deposition medium, wet-dry cycles with water and wet-dry
cycles with deposition medium.

AVG R

AVG N

AVG C

AVG NC

AVG
NCS3%

AVG
NCS5%

0.0007

K (m/s)

7E-05

7E-06

In water
In medium
WD+water
WD+medium

7E-07

7E-08

Fig. 2.12 Water permeability coefficient of samples in different curing conditions

90
80

Healing (%)

70
60

95% RH
In water
In medium
WD+water
WD+medium

50
40
30
20
10
0
R

NC

NCS3%

NC5%

Fig. 2.13 Healing ratio of samples in various curing conditions

Observation were made 56 days after cracking, it was concluded that specimen in series NCS3%
showed maximum crack healing while those of NCS5% gave lowest water permeability results
because of the reason that greater amount of inert microencapsulation caused water-proofing

15

effect and didnt allow contact of bacteria with water, hence decreasing self-healing ration and
permeability. While observing effect of incubation techniques it was evident that wet dry cycle
with water was the most effective technique as during wet cycle specimen absorbed water and
during dry cycle it got more oxygen from atmosphere.
In addition to type of bacteria, selection of carrier is also a matter of great significance. When it
comes to strength of concrete and viability of bacteria, carrier material is an important factor in
the manufacturing of self-healing concrete Therefore, studies related to the carrier materials are
also presented.
Gadea et al. (2010) studied the use of polyurethane foam wastes (PFW) in making lightweight
cement based mortar. During his research he grounded the PFW to the size less them 4 mm and
gradually replaced fine aggregate with grounded PFW. Two types of cement were used in this
study i.e. Cem I 42.5 R and Cem IV 42.5 N. mechanical properties including workability,
permeability and strength were checked for each replacement level. It was found that inclusion
of PFW increases the workability of mix and at the replacement level of 100% it increases the
workability of mix by 120 min. on the other hand, effect of PFW on the flexural and compressive
strength of mix at various checking days was devastating.
9
CEM I 42.5 R

Flexural strength (MPa)

CEM IV 42.5 N

7
6
5
4
3
2
1
0
0

25
50
75
Sand replaced by polyurethane foam waste (%)

100

Fig. 2.14 Flexural strength at various replacement levels of polyurethane foam wastes

16

Sixuan (2012) studied the possibility of using graphite nano platelets (GNP) in cement based
construction materials. During his study, GNP particles were added in the cement mix and both
mechanical and electrical properties of cement mix were studied. To study the compressive and
flexural strength of cement mix two different kind of samples were made. 50 x 50 x 50 mm
cubes were made to study the compressive strength of mortar and 40 x 40 x 160 mm prisms were
made to study the flexural properties. During this experimental work it was found that increasing
the amount of GNP in mortar has a better effect on the compressive and flexural properties of
mortar. Three different percentages of 0.5, 1 and 1.5% were used. It was seen that mix with 1.5
% GNP had maximum compressive strength where mix with 1% GNP had maximum flexural
strength and flexural strength started to decrease when percentage was increased to 1.5%.
70
Compressive stength

Strength (MPa)

60

Flexural strength

50
40
30
20
10
0
0

0.50%

1.00%

1.50%

GNP/Cement (%)

Fig. 2.15 Effect of GNP on strength of cement based mortar

This difference in healing ratio, threshold crack healing and difference in compressive strength in
various studies can be attributed to a number of factors such as type of bacteria, their varying
concentrations, various curing procedures, nutrient compounds and material used for
incorporation of bacteria. To identify most practical self-healing techniques in concrete, there is
a need to determine the most effective bacteria along with its optimum concentration, a host
precursor and activation process.
It is evident from the researches presented here that there is still not enough data to determine the
most effective bacteria and its optimum concentration. As we compare the results of various
17

researches there is a considerable difference in self-healing capacity displayed by the procedures.

These results are for the cracks in specimens with B.Sphaericus as healing bacteria, incorporated
in mix using various techniques. Comparison of the results shows a considerable variation in
permeability of self-healing concrete. Technique of incorporating bacteria with microencapsulation exhibits better healing as compared to others, however, micro-encapsulation is a
comparatively difficult process and needs extra care. Considering the variation displayed by
different techniques, it is hard to recognize an optimum carrier with better incorporation
techniques.
Owing to the low tensile strength of concrete, it is desired that introduction of carrier compounds
in self-healing concrete give beneficial effects by providing protective cover to bacteria as well
as enhancing the flexural tensile strength of concrete. Fig. 2.16 shows the comparison of flexural
strength provided through polyurethane (PU), light weight aggregate (LWA) and graphite nano
platelets (GNP). It can be seen that PU and LWA cause decrease in the flexural strength of
mortars instead of increasing it. GNP on the other hand provides better flexural strength to the
mortar and at 1% addition of GNP maximum flexural strength was observed. It is evident, out of
three presented methods only GNP increases the flexural strength of concrete, therefore GNP is
considered helpful in reducing crack formation in concrete.
15

Flexural Strength ( MPa)

10
5
0
-5
-10

Control

-15

Bacteria Incorporated
Percentage Difference

-20

LWA

PU

GNP

Fig. 2.16 Comparison of flexural strength in PU, GNP and LWA incorporated concrete
Not only the choice of carrier compound affects the self-healing capacity but choice of bacteria
selection also plays a great role in the self-healing process. Results on compressive strength by
18

Ramachandran et al. (2001) show that using bacteria, B.Pasteurii, 28 days strength of concrete
increased by 18% at concentration of 7.6 x 103 cells/cm3, whereas the research work done by
Ghosh et al. (2005) shows that bacteria, Shewanella, at the concentration of 105cells/cm3 results
in 25% increase in 28 day strength than the control samples which is greater as compared to 18%
increase as reported by Ramachandaran and 2% increase in strength as measured in case of
E.Coli. In the case of B.Pseudofirmus, it can be see that at a concentration of 6 x 108 cells/cm3
results in a 10% decrease in the strength of mortar. in addition to that, research work done by
Wang et al. (2012) shows that compressive strength of mortar decreased with the increasing
replacement level of B.Sphearicus and show an approximately 35% decrease in 28 day
compressive strength at the replacement level of 5%. Fig. 2.17 shows a comparison of effect of
different bacteria on strength of mortars.
Compressive strength variation
(%)

30
20
10
0

-10
-20
-30
-40
Bacillus
Pastuerii

Shewanella

E.Coli

In Concrete In Concrete In Concrete

Bacillus
Bacillus
Pseudofirmus Sphearicus
In Mortar

In Mortar

Fig. 2.17 Effect of various bacteria on compressive strength of self-healing concrete

These results depict that there is still uncertainty about appropriate bacteria and their optimum
concentration that can have a better impact on compressive strength of concrete. Therefore, work
is required to study the impact of new bacteria and incorporation techniques on the compressive
strength of concrete to determine optimum conditions.

3. Summary
A review of previous studies on self-healing techniques has been presented. It is seen that quite
limited type of bacteria and carrier compounds have been used in the previous studies. The
19

purpose of this research is to collect sufficient data on the efficiency of Bacillus Subtilis and use
of graphite nano platelets to check the possibility of an alternate bacteria and carrier compound
for self-healing studies that can lead to most conducive and favourable outcome in self-healing
of concrete.

20

CHAPTER 3
EXPERIMENTAL PROGRAM
1. General
In recent years, several studies have brought the attention towards the importance and
significance of the emerging technique of bio-influenced self-healing concrete. These studies
were, however, limited to a small group of bacteria and protective carrier compounds. Therefore,
there is a need to carry out in-depth study on new alternatives in the form of both bacteria and
protective carrier compounds. Evaluating the best suited bacteria and ideal protective carrier
materials can be helpful in achieving an optimum self-healing process in bio-influenced
concrete. Compressive strength tests and pre-cracking techniques were applied to concrete
specimen incorporated with precursor materials and carrier compounds containing bacteria.
Description of specimen for these tests is provided in the Table 3.1.
Table 3.1 Description of test specimens
Sample dimensions for compressive
strength tests

Specimen

Sample dimensions for pre-cracking

Dia (in)

Height (in)

Dia (in)

Height (in)

Mix 1

12

Mix 2

12

Mix 3

12

Mix 4

12

Specimen names indicate the techniques and methods used for the incorporation of bacteria in
them. Control specimens containing neither bacteria nor organic pre-cursor compound are
denoted by Mix 1, whereas, Mix 2 shows that bacteria were incorporated in those specimens
directly without the use of any protective carrier compounds. In the same way, those
incorporated with bacteria by the use of light weight aggregates (LWA) as protective carrier are
represented by Mix 3 and specimen containing graphite nano platelets as a mean of bacteria
inoculation are termed as Mix 4.
21

The tests for compressive strength were conducted according to criteria defined in ASTM C-39
(revised 2014) and ASTM 2809 standard was followed during the process of scanning electron
microscopy. Crack widths were measured by the use of crack width measuring microscope with
a least count of 0.02 mm. This chapter describes the experimental program including testing
procedures. Details about material properties, sample preparation and testing methodology have
also been included in this chapter.

2. Materials
The important feature of this study is the comparison of various techniques of bacterial
incorporation in concrete. It is being used to compare the results achieved by use of both GNP
and LWA and determine the better choice. The constituent materials used in this study were
tested conferring to their relevant standards and to ensure consistency in their supply, periodical
quality assurance tests were also carried out. Tests conducted on different materials used in this
study are described below.

2.1

Micro-organism

In order to be used as a healing compound in the concrete samples, the micro-organism had to
possess certain features. It must be able to withstand alkaline atmosphere of concrete in order to
carryout production of calcium carbonate, it should produce copious amount of calcium
carbonate without being affected by calcium ion concentration, it must be able to withstand high
pressure and should be oxygen brilliant to consume much oxygen and minimize corrosion of
steel (Gupta et al., 2013; Jonkers et al., 2010; Rao et al., 2013). Bacteria namely, Bacillus
Subtilis, was selected in this study as it fulfilled the necessary criteria for survival in harsh
environment. It is gram-positive bacteria having an ability to form spores when subjected to
unfavourable conditions. This spore formation provides its protection against high mechanical
pressure and alkaline environment, making it ideal selection. Members of genus bacillus can
produce spores which can lay dormant for over 200 years (Schlegel, 1993).
Bacterial solution (bacteria in nutrition bath), specially prepared and treated to ensue spore
formation in controlled microbiology laboratory, in usually used to introduce these bacteria in
different incorporation techniques in concrete. Fig. 3.1 shows the bacteria solution containing
spores of B.Subtilis bacteria. The quantity of solution required in the mix was calculated on the
basis of concentration found by optical density test using a spectrophotometer. For this purpose,
22

medium in which bacteria was growing in was selected as blank. This blank solution was used as
a reference, on the basis of which optical density of bacterial solution was measured. Initially 0.5
ml of blank solution was placed in spectrophotometer and 600 nm wavelength was selected.
After the machine had read the blank solution, 0.5 ml of bacterial solution was placed in
spectrophotometer and wavelength of 600 nm was selected to carry out the optical density test.
On the basis of this test, concentration of bacteria in the solution was measured using the
expression Y=8.59 x 107 X1.3627 (Ramachandran et al., 2001). Where Y is the bacterial
concentration per mL and X is the reading at OD600. With spectrophotometer, the bacterial
concentration was found to be 2.8x108 cells/ml. Based on these results, spore concentration in
samples was kept equal to 3x108 cells/cm3 of concrete mixture.

Fig. 3.1 Bacteria solution in a cylinder

2.2

Graphite nano platelets

Studying the effect of graphite nano platelets (GNP) as carrier material and its comparison with
other carrier compounds is the core objective of this research. Therefore, evaluating the
properties of GNP is of prime importance. Physical appearance of GNP is shown in Fig. 3.2.

23

(a) Graphite nano platelets

(b) Microstructture of raw graphite

nanoplatelets

Fig. 3.2 Graphite nano platelets

The material properties of GNP provided by supplier are given in the Table 3.2
Table 3.2 Properties of GNP
Properties of GNP
Grade

Nano 25

Primary particle size

100nm

Composition

99 % carbon

Surface Area

180 m2/g

Lamella thickness index (LTI)

14-15

Absorption tests were also conducted on GNP to find their maximum absorption capacity. This
test helped to determine the amount of GNP required as a carrier compound for bacteria.
Graphite nano platelets were submerged in water for 24 h and were filtered by the help of filter
paper. After measuring the saturated weight of GNP, it was oven dried and weighted again to
calculate water absorption. On the basis of this test water absorption of GNP was found to be
62%

2.3 Light weight aggregate

Using light weight aggregates (LWA) was studied as a carrier material and was compared with
GNP to evaluate the optimum method of bacteria transfer in the mix. Fig. 3.3 shows the LWA
used in this study for the incorporation of bacterial spores.
24

Fig. 3.3 Light weight aggregates

Like GNP, water absorption test was also conducted on LWA to determine the exact amount of
LWA to be used. To evaluate the water absorption in light weight aggregate 500 gm of LWA
were submerged in water to make it saturated. After 24 h they were surface dried and their
saturated weight was measured. These aggregates were later placed in oven and were dried for
24 h and dry weight was measured. On basis of this test water absorption of LWA was found to
be 16.6%

3. Mix proportion
Four different types of mix were used for the study. The mix proportion for these four different
categories of specimens contained ordinary portland cement (OPC) type I conforming to
ASTM C 150-07 as 370 kg/m3, fine aggregate as 840 kg/m3, coarse aggregate as 990 kg/m3 and
calcium lactate of 18 kg/m3 with a water to cement ratio of 0.4 for all the mixes for concrete. The
mix was designed for a compressive strength of 4000 psi. Sikament - 520 set-retarding
admixture was used as 1% by weight of cement for producing free-flowing concrete in hot
climates. ASTM C 191-11 and ASTM C 187-11 codes were conformed for normal consistency
test and initial and final setting time respectively for cement. Control specimens were named
Mix 1 in which no bacterial spore specimens were added. In Mix 2 specimens, bacteria were
incorporated directly by mixing the bacterial solution in water during mixing of concrete without
use of any protective carrier compound. In the same way, those incorporated with bacteria by use
of LWA as protective carrier were labelled as Mix 3. In order to incorporate bacteria, LWA
were kept soaked in bacterial solution for 24 h till they were saturated prior to their mixing in
25

concrete. Specimens containing GNP as a mean of bacteria introduction were termed as Mix 4.
GNP was also soaked with bacterial solution before mixing in concrete. However, in order to
ensue uniform distribution of GNP in concrete, superplasticizer (Sikament - 520) was added to
GNP soaked bacterial solution. Addition of superplasticizer in GNP prior to mixing in concrete
ensures uniform distribution of GNP particles throughout the concrete mix (Sixuan, 2012).
The mixing procedure and time were kept constant for all the concrete mixes investigated and
details of mixing proportions for different batches of concrete are presented in Table 3.3.
Table 3.3 Mix design of different sets of specimens
Specimens

Mix 1

Mix 2

Mix 3

Mix 4

Cement

kg/m3

370

370

370

370

Fine Aggregate

kg/m3

840

840

840

840

Coarse Aggregate

kg/m3

990

990

990

990

0.4

0.4

0.4

0.4

kg/m

18

18

18

18

liter/m3

6.33

6.33

6.33

None

Direct

By LWA

By GNP

Water cement Ratio

Super Plasticizer
Calcium Lactate
Bacteria with Spore
concentration
(2.8 x 108 cells/ml)
Bacteria incorporation
technique

(%)
3

4. Test specimens
Specimens were removed from moulds after 24 h of casting and were placed to be cured in
controlled conditions. For all mix types, samples for two different dimensions were made. For
pre-cracking specimens of 150 mm dia and 100 mm height were prepared and five specimens
pre-cracked at 3,7,14 and 28 days for each mix were studied for healing measurements. For the
compressive strength tests cylindrical specimen of diameter 150 mm and height of 300 mm were
prepared and an average of three specimens were utilized to determine 3,7,14 and 28 days
compressive strength. Fig. 3.6 shows prepared specimens for different tests to study the material
properties. Test specimens were made in accordance to ASTM C 39. Proper tamping was carried
out to ensure the compaction of specimens. Moreover, samples were also subjected to scanning
electron microscope (SEM) analysis to monitor microstructural changes due to mineral
26

formation. In addition, samples of mineral produced during the process of self-healing was taken
and was subjected to X-Ray Diffraction analysis (XRD) to identify its chemical composition.

Casting of samples for pre-cracking

Casting of samples for compressive

strength measurement

Fig. 3.4 Casting of samples

5. Test procedure
The tests for compressive strength were performed on 3, 7, 14 and 28 days cured samples and an
average of three samples was taken. At the appropriate age the specimens were removed from
water and surface water was wiped off. In order to ensure uniform distribution of load on the
face of sample, proper capping was done prior to placing sample in the compression machine as
shown in Fig. 3.5. Sample was placed in testing machine and load was applied on permissible
rate.
However, for pre-cracking of samples, samples were monitored carefully under controlled
compressive load. Application of load was stopped immediately after the development of visible
cracks. The crack widths were measured at different points on the specimens and the cracks with
a width around 1 mm were selected and marked for further observation of self-healing. Initial
crack width was measured at the time of crack development by the help of crack width
measuring microscope, accurate up to 0.02 mm.

27

Capping of samples

Compression testing

Cylinder after compresssive

machine

strength test

Fig. 3.5 Capping of cylinders, compression machine and cylinders after compressive
strength tests
Fig. 3.6 shows the crack measuring microscope. Cracks were marked to ensure that crack widths
are measured at the same location every time as shown in Fig. 3.7. After measuring the initial
crack width, the pre-cracked specimens were continued to cure under controlled curing
conditions. At appropriate age of 3,7,14 and 28 days, these pre-cracked specimens were taken
out and crack width was measured again by using crack width measuring microscope. However,
during this study visual inspection was carried out for this purpose as shown in Fig. 3.8 and
difference between crack widths measured initially and on later stages was calculated as the
measure of self-healing efficiency of concrete. An average of three tests on specimens was taken
for compressive strength, while for self-healing measurement an average of five test specimens
was accounted.

28

(a) Crack measuring microscope

(b) Crack width measurement by crack

measuring microscope

Fig. 3.6 Crack measuring microscope and measurement of crack by it

Fig. 3.7 Marking of cracks on pre-cracked specimens to ensure that multiple reading are
taken from the same point

29

Day 3

Day 7

Unhealed Crack
Formation of CaCO3

Day 14

Day 28

Day 3
Formation of CaCO3
Unhealed part
of crack

Completely healed crack

Fig. 3.8 Closer view of cracks showing self-healing process with calcium carbonate formation
In addition to these tests, samples were collected form the cracked samples at the age of 7 and 28
days after pre-cracking and were subjected to scanning electron microscopy (SEM).
Results from compressive strength tests, visual inspection of crack healing, SEM and XRD were
recorded. These results were later analyzed and compared to reach the conclusion regarding
comparison of techniques. Comparisons of these results along with related discussion are
presented in chapter 4.

30

CHAPTER 4
RESULTS AND DISCUSSIONS
1. General
Results from previously mentioned tests are presented and discussed here to determine the
efficiency of self-healing process of all mixes. These results include the crack width
measurements, visual inspection of cracks, compressive strength of self-healing concrete samples
and micro structural study through SEM and mineral composition of healing compounds through
XRD analysis. Furthermore, relationships of self-healing at different curing days and
compressive strength with self-healing techniques are elaborated.

1.1 Self-healing analysis

Pre-cracked specimens were monitored at specified time of 3,7,14 and 28 days to determine the
efficiency of self-healing process by use of crack measuring microscope. These cracks showed
significant self-healing specially in specimens of Mix 2, Mix 3 and Mix 4. Production of calcium
carbonate crystals (CaCO3) in significant amount was observed in these cracks as shown in Fig.
4.1. The healing compound, CaCO3 crystals, was the main factor in reducing the cracks width in
bacteria incorporated specimens. This formation of healing compound, as a result of bacterial
conversion of calcium lactate into calcium carbonate, was also observed and discussed by
Jonkers (2010).

Fig. 4.1 Crack width measurement by crack measuring microscope

31

The measure of healing is obtained in millimetres as the difference of initial crack width and
healed crack width in different predefined times. Fig. 4.2 illustrates the efficiency of crack
healing of each mix as a function of time. While observing specimens of all incorporated
techniques pre-cracked on 3 days of curing, healing of cracks was prominent especially after 7
days of curing. However, as shown in Fig. 4.2 healing efficiency of Mix 4 samples with GNP as
a carrier compound show the maximum healing as a function of time on all crack measuring
days. This increase in healing in Mix 4 is due to the particle size of GNP. The small size of GNP
enables it to act like a filler material (Sixuan, 2012) and assures its uniform distribution
throughout the mixture. GNP particles are saturated with bacteria medium, resulting in uniform
distribution of bacteria through colloidal dispersion in the mix and ensuring the availability of
bacteria medium at the crack site for formation of healing compound. These self-healing bacteria
when come in contact with water, ingressed through cracks, gets activated and carry out the
conversion of calcium lactate into water insoluble calcium carbonate which results in crack
healing. Mix 4 specimens are followed by specimens containing LWA as a carrier material.
While LWA has particle size greater than that of GNP, therefore it becomes difficult to ensure
the distribution of LWA in the mix as uniformly as GNP. Bigger size of LWA makes it harder
for LWA to get in the inter particle spaces where GNP can penetrate easily owing to its nano
sized particles. This feature of LWA hinders the equal and even distribution of bacteria in the
mix hence decreasing the efficiency of self-healing process in concrete. Fig. 4.2 illustrates that
all the samples incorporating bacteria had showed improvement in healing results as compared to
control samples. Controlled concrete samples, without any bacteria, showed small crack healing
as well, which can be credited to a number of reasons. Some cement particles are not completely
hydrated during the initial mixing process and at early age. These particles go through continued
hydration resulting in production of expansive hydration products which eventually lead to
healing of cracks. Another factor for crack healing in controlled samples is the carbonation
process of calcium hydroxide. This carbonation process leads to conversion of calcium
hydroxide to calcium carbonate.(Homma et al., 2009; Jonkers and Schlangen, 2009; Wiktor and
Jonkers (2011)).
Calcium carbonate production in controlled specimens is mainly dependent on the availability of
carbon dioxide (CO2) dissolved in the ingressed water. However, the amount of carbon dioxide
dissolved in water is limited, which in turn limits the production of calcium carbonate due to
32

carbonation process. Furthermore, portlandite (Ca(OH)2) is soluble in water therefore, the

ingressed water not only provides carbon dioxide for the carbonation process but it also dissolves
the precipitated calcium hydroxide formed in controlled samples, leaving less calcium hydroxide
in concrete samples for conversion into calcium carbonate.
According to Ter Heide (2005) healing in controlled samples can also be attributed to the
swelling of cement matrix.
Contrary to that, the process of self-healing in bio concrete is different due to the presence of two
component healing system i.e. calcium lactate and self-healing bacteria. After coming in contact
with the water ingressed through cracks dormant bacterial spores get activated. As a result of
bacterial activity, calcium lactate present in bio concrete is converted directly into calcium
carbonate, which is insoluble in water. This bacterial activity also results in the production of
CO2 within the specimens, as a by-product of metabolic reactions, hence ensuring the availability
of more carbon dioxide for the process of calcium hydroxide carbonation leading to production
of calcium carbonate. Both of these process take place simultaneously within the bio concrete
samples resulting in two fold production of healing compound (Schlangen et al., 2010).
0.9
Mix 1

Healing (mm)

0.8

Mix 2

0.7

Mix 3

0.6

Mix 4

0.5
0.4
0.3
0.2
0.1
0

7
Healing time (days)

14

28

Fig. 4.2 Crack healing in specimens pre-cracked at 3 days

Crack healing in specimens pre-cracked on 7 days of curing show similar trend observed in
specimens pre-cracked at 3 days as shown in Fig. 4.3. Fig. shows that Mix 4 specimens, having
33

GNP as carrier compound, show much more healing as compared to all other techniques. As
previously discussed, this efficient behaviour of GNP is due to its small size which ensures
thorough and even distribution of particles carrying bacteria in the mix. This small size allows
GNP particles to enter the inter-particle spaces and make the availability of bacteria possible for
the process of self-healing. LWA, being bigger in size as compared to GNP, cannot penetrate in
the micro sized gaps which limits the capacity of LWA to evenly distribute self-healing bacteria
in concrete matrix. The maximum healing of cracks observed in Mix 4 after 28 days was 0.81
mm. Whereas, crack healing of 0.61 mm was observed in Mix 3 specimens, comprising of LWA.
Mix 2 samples, prepared by direct introduction of bacteria in the concrete mix, showed healing
of 0.37 mm. However, comparison of results achieved from 3 and 7 days pre-cracked specimens
indicates that there has been a slight decrease in self-healing observed in specimens of all mix
types. This decrease in self-healing is because of an overall reduction in further production of
hydrated products in concrete as significant amount of cement is already hydrated. Another
reason for this overall reduction in bio-concrete specimens is due to slight decrease in viability of
bacterial survival in concrete as development of denser micro-structure in concrete is started.
Mix 3 and Mix 4 are least affected by denser micro-structure at this stage, because at this stage
pressure exerted by microstructure development is not significant and carrier compounds are able
to provide better cover to bacteria against the exerted pressure.
0.9
0.8

Mix 1
Mix 2

Healing (mm)

0.7
0.6

Mix 3
Mix 4

0.5
0.4
0.3
0.2
0.1
0
3

14

28

Healing time (days)

Fig. 4.3 Crack healing in specimens pre-cracked at 7 days

34

Healing observed in specimens pre-cracked after 14 days of curing is expressed in the Fig. 4.4.
Mix 3 specimens show maximum healing in samples pre-cracked at 14 days of curing. Mix 4,
which displayed maximum healing in samples pre-cracked at 3 and 7 days of curing, proved less
efficient than Mix 3 when pre-cracked at 14 days of curing.
0.7
Mix 1
0.6

Healing (mm)

0.5

Mix 2
Mix 3
Mix 4

0.4
0.3
0.2
0.1
0
3

14

28

Healing time (days)

Fig. 4.4 Crack healing in specimens pre-cracked at 14 days

This change in behaviour of Mix 4 can be due to continued development of dense micro
structure resulted by continued hydration reactions in concrete. This dense micro structure in
concrete creates a pressure on the carrier compounds containing incorporated bacteria. This
pressure was not resulting in a significant affect over the viability of bacteria survival in
specimens pre-cracked at 7 days. However in specimens pre-cracked at 14 days of curing this
pressure has been increased enough to have a considerable effect on the self-healing capacity of
all specimens. The effect of micro-structural pressure is most significant in specimens of Mix 2
and Mix 4. On the other hand, LWA is seen to be least affected by this pressure. GNP is weak
when subjected to multi axial loading (Sixuan, 2012), hence, it is unable to provide better cover
to bacteria as compared to that provided by LWA. This leads to elimination and annihilation of
bacteria in Mix 4, therefore healing process observed in Mix 4 was decreased than it was in
samples pre-cracked at 3 and 7 days. This decline in self-healing can be attributed to
underdeveloped microstructure which is not fully matured till only 7 days of casting and
becomes more compact and mature at 14 days. The change in self-healing process can also be
35

observed in the Mix 2 specimens, with directly incorporated bacteria, with the value of 14 days
healing decreased from 0.37 mm to 0.21 mm. This decreases in crack healing of Mix 2 samples
is due to decrease in the viability of bacteria survival in concrete under the pressure applied
during the mixing phase and that developed due to formation of dense micro structure (Jonkers et
al., 2010).
Jonkers and Schlangen (2009) attributed this decrease in self healing activity to disintegration of
calcium lactate in the concrete mix. However, small amount of calcium lactate in his research
was used (1% of cement weight) and in this research 4.86% of cement weight has been used to
ensure enough availability of calcium lactate to the bacteria. Furthermore, this drastic decrease in
bacterial activity was only observed in Mix 2 specimens, which shows that this decrease is the
result of elimination of bacteria and not because of calcium lactate disintegration..
Fig. 4.5 depicts the results obtained through measurements of self-healing in samples precracked after 28 days of curing. It can be seen that Mix 3, with LWA, is showing maximum
healing of 0.52 mm, higher than all other mixes. Mix 4 specimens, with GNP, shows much less
healing than it showed at 3 and 7 days of pre-cracking. The healing exhibited by Mix 4
specimens was 0.38 mm, which is higher than healing of 0.15 mm showed by Mix 2 specimens.
0.6
Mix 1
0.5

Mix 2

Healing (mm)

Mix 3
0.4

Mix 4

0.3
0.2
0.1
0
3

14

28

Healing time (days)

Fig. 4.5 Crack healing in specimens pre-cracked at 28 days

36

This shows that Mix 4 still provides a significant improvement in healing with GNP incorporated
bacteria but this improvement in healing is lower than that exhibited by Mix 3. This reduction in
healing of Mix 4 specimens at 28 day pre-cracking can again be attributed to the dense
microstructure formed in the concrete after 28 days of curing similar to samples pre-cracked at
14 days of curing.

1.2 Microstructure analysis

In addition to the results achieved by visual inspection of concrete samples, specimens of all four
mixes were subjected to scanning electron microscopy (SEM) analysis to study changes in
concrete microstructure due to self-healing. For comparison of self-healing process SEM
analysis was conducted at 7 and 28 days of healing.
Production of calcium carbonate based crystals were the main focus of this study as it expresses
the crack healing efficiency of respective mix. Calcium carbonate crystals are developed in three
different forms which are named calcite, aragonite and vaterite (Rao et al., 2013). Out of the
three, calcite is most stable form of calcium carbonate.
Fig. 4.6 shows SEM images of all four mixes at 7 days pre-cracked specimens with 2 m
resolution. Fig. shows the development of calcite crystals which are orthorhombic in nature (De
Yoreo and Vekilov, 2003). Mix 4 containing GNP as carrier compound showed maximum
calcium carbonate (CaCO3) formation as compared to mixes with other techniques.
This CaCO3 formation results from presence of two component, bacteria and calcium lactate,
based healing system. Although CaCO3 is also formed in controlled samples, however presence
of bacteria along with calcium lactate catalysis the production of CaCO3 crystals. CaCO3 crystals
formation in higher quantities and similarities in shape is identical and conform to those reported
by Jonkers et al. (2010) and Wang et al. (2012). The chemical process of calcium carbonate
formation by bacterial activity is presented in equation given below.
CaC6H10O6 + 6O2 CaCO3+5CO2+5H2O

(1)

As stated earlier that production of CaCO3 is not only limited to concrete with bacteria
incorporated in them. The presence of CaCO3 is also evident in controlled concrete specimens.
However, the process of CaCO3 crystals formation in controlled specimens is quite different to
that in bacteria incorporated specimens. Formation of CaCO3 in Mix 1 specimens is due to the
37

carbonation of calcium hydroxide, as given in equation 2, which is one of the major hydration
products of cement.
Ca(OH)2 + CO2 CaCO3+ H2O

(2)

CaCO3 Crystals

C-S-H Gel

A : Mix 1

B : Mix 2

CaCO3 Crystals

CaCO3 Crystals

C : Mix 3

D : Mix 4

Fig. 4.6 Scanning electron microscope analysis of 7 days pre-cracked samples

However, this production of CaCO3 crystals in Mix 1 due to carbonation process is very slow as
compared to those produced by bacterial activity. In addition, as calcium carbonate production in
controlled specimens is due to the availability of carbon dioxide (CO2) dissolved in the ingressed
water; therefore, less amount of CO2 is available for carbonation process. Furthermore, as
portlandite (Ca(OH)2) is soluble in water so whenever it comes in contact with ingressed water it
gets mixed in it, leaving less calcium hydroxide on the contact surface to convert in CaCO3. On
the other hand, in bio concrete, the process is different due to the presence of calcium lactate and
38

bacteria. Bacteria converts calcium lactate directly into calcium carbonate which is insoluble in
water and as result of this metabolic reaction CO2 is produced which reacts with calcium
hydroxide on spot and does not let it wash away. Hence, producing more calcium carbonate
(Schlangen et al., 2010).
Fig. 4.7 shows SEM analysis of 28 days pre-cracked specimens. It can be seen that calcium
carbonate crystal formation is higher in Mix 3, observed in specimens pre-cracked at 28 days
compared to Mix 2. The amount of CaCO3 in Mix 2 seems even less as compared to those
produced in samples pre-cracked at 7 days of curing. Curing for 28 days improves the
development of microstructure within the concrete samples, making it denser. This results in
decreasing the pore sizes and application of pressure on self-healing bacteria. This leads to
elimination of bacteria, leaving less number of bacteria in specimens for the production of
calcium carbonate.
Fig. 4.6 shows that Mix 4 displayed much more crystal formation in 7 days pre-cracked samples
as compared to Mix 3. However, as shown in Fig. 4.7, GNP is no longer able to provide effective
cover to bacteria and therefore, calcium carbonate crystal formation in Mix 4 is reduced
significantly compare to CaCO3 crystal produced in Mix 3. These results depict that in samples
pre-cracked at 28 days of curing; LWA provides the best cover to bacteria. As described by
Sixuan (2012), GNP are weak when it comes to multi-axial load application and does not provide
better cover to bacteria. Thus, with the increase in completion of hydration reaction and decrease
of pore size, healing efficiency of Mix 4 samples decreases. However, LWA provides cover
during the mixing phase and provides better protection to spores in the samples as it provides
resistance against the pressure developed in samples due to microstructure development. The
variation in CaCO3 formation with and without carrier compound conforms the trends seen in the
results, in the study carried out by Wiktor and Jonkers (2011). The crystals formation observed
in SEM images are also similar to those observed and presented by Wang et al. (2012) and Wang
et al. (2014) which confirms the formation of calcium carbonate with similar crystalline
structure.

39

Ettringite

CaCO3 Crystals

A : Mix 1

B : Mix 2

CaCO3 Crystals
CaCO3 Crystals

D : Mix 4

C : Mix 3

Fig. 4.7 Scanning electron microscope (SEM) analysis of 28 days pre-cracked samples

1.3 X- Ray diffraction analysis

For better understanding of self-healing process and to verify the formation of calcium carbonate
in the samples, the healing compounds developed in cracks were subjected, to XRD analysis. In
order to get the sample, the healing product formed inside the cracks was scratched with great
care and was placed in the XRD apparatus. Copper (Cu) was selected as a X-ray target because it
can be kept cool easily, due to its high thermal conductivity, and which produces strong K and
K lines. The readings were recorded at a wavelength of 1.54 and different representative
peaks were obtained as shown in Fig. 4.8. It can be seen from the Fig. that no sharp needle like
peaks were obtained during XRD. This is due to the reason that sample was scratched from the
crack surface and contained a mixture of compounds from concrete surface as well.
40

Fig. 4.8 XRD analysis of healing compound produced in the cracks.

Bacillus Subtilis is calcite forming bacteria (Rao et al., 2013), therefore, the results of XRD were
compared with the reference cards of calcite. A highest peak was obtained at the 2theta (2)
value of 29.2070o which is quite close to 2 of 29.455o of pure calcite as observed by
(Herrington, 1927). The slight difference in 2 value can be due to the impurities in the powder
resulted from the scratching off process. This shows that the material produced in the cracks is
calcium carbonate in nature and is in harmony with the results obtained from previous studies.

1.4 Compressive strength analysis

Measured compressive strength of self-healing specimens is presented in Fig. 4.9. It can be seen
that all bacterial incorporation techniques result in increased compressive strength of the mix.

41

Compressive strength (MPa)

35

30

Mix 1
Mix 2
Mix 3
Mix 4

25
20
15
10
3

7
14
Curing time (days)

28

Fig. 4.9 Compressive strength development with different bacteria incorporation techniques
Samples having LWA as a carrier compound for bacteria incorporation showed maximum
strength of 29.43 MPa and improvement of 12 % in compressive strength as compared to
controlled concrete specimens. The increase in compressive strength are in accordance with the
results as recognized in the study in self-healing carried out by Sierra-Beltran and Jonkers (2012)
and confirms that self-healing is a cause of increase in compressive strength. This increase in
compressive strength can be attributed to smaller size of LWA in comparison to regular sized
coarse aggregates. This allowed better packing and compaction of concrete matrix around them
which gave these specimens much higher strength than controlled specimens.
Specimens containing GNP showed an increase of 9.8% in compressive strength. This
improvement in compressive strength can be attributed to the addition of GNP. GNP being a
nanosized material acts like a filler material with even and uniform suspension in the mix. Small
size of GNP also decreases the formation of weak interfacial transition zone (ITZ) in concrete by
allowing filling of porous and crystalline microstructure within ITZ. Decrease in ITZ makes the
mortar matrix denser and more compact resulting in higher compressive strength. GNP particles
also act as crack arrestors and block and divert crack formation and propagation (Sixuan, 2012).
GNP therefore, acts in many ways to enhance the compressive strength of concrete.
Direct incorporation of bacteria also showed an increase in compressive strength of concrete.
This improvement is because of the presence of calcite producing bacteria in the mix. These
42

calcium carbonate continuously manufactured by the bacteria and calcium lactate provided as
organic precursor makes the internal structure of concrete more compact, therefore, results in
increase of compressive strength. This improvement seen by the direct introduction of bacteria is
in consistence with the results achieved by Ghosh et al. (2005). However, after careful
comparison of the results achieved by Ramachandaran (Ramachandran et al., 2001) by direct
introduction of bacteria, it is evident that there is no difference in strength by introduction of
Bacillus Pasteurii at a rate of 7.2 x 107 cell/cm3. This shows that as far as compressive strength is
related B.Subtilis is a better choice as compared to B.Pasteurii as its addition significantly
improves the compressive strength of concrete.

43

CHAPTER 5
CONCLUSIONS AND RECOMMENDATIONS
1. General
The aim of this study is to compare the efficiency of different self-healing techniques in terms of
their crack healing capacity. Efficiency of these techniques was studied in terms of their effects
on compressive strength, micro structure, production of calcium carbonate and healing of cracks.
SEM analysis was carried out to observe changes in micro-structure because of various
techniques. In addition, healing compound observed in specimens was subjected to XRD in order
to determine its chemical composition. Results required from experimental work were used to
develop the relationships of observed crack healing and compressive strength with bacteria
incorporation techniques.

2. Conclusions
Based on the results achieved during this study following conclusions are drawn:

Specimens incorporated with graphite nanoplatelets (GNP) as carrier compound displayed

uniform distribution and protection of bacteria at samples pre-cracked at early age of 3 and 7
days, resulting in maximum crack healing efficiency. However, when pre-cracked at later
days, such specimens presented a significant decrease in healing of cracks.

Although specimens incorporated with lightweight aggregate (LWA) as carrier compound,

were not as efficient as GNP at early age pre-cracked specimens, they showed consistency in
their crack healing efficiency in specimens pre-cracked at later days.

Specimens incorporated directly with bacteria did not show any effects in crack healing of
concrete.

Compressive strength trends of all mixes suggest that, addition of bacteria Bacillus Subtilis
resulted in slight increase in compressive strength, irrespective of the incorporation
technique, with significant improvement through LWA technique.

3. Recommendations
While this research has provided significant understanding about self-healing mechanism and the
effect of newly introduced carrier compound on crack healing capacity, further research is
44

required to completely characterize the self-healing mechanism. Given below are some of the
recommendations for further research in this area.

There is still a need of further research in order to determine the optimum concentration of
bacteria to maximize the production of biologically produced CaCO3 and increase
compressive strength.

Further study is required for the detailed understanding of activation process of bacteria and
to find the methods to control them.

Experiments should be carried out to determine the durability of bacteria based self-healing
concrete.

45

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